LOCUS       sequence-264203-        4347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4347)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 4347)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221_SLC6A1
FEATURES             Location/Qualifiers
     source          1..4347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2472..2571)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2588..2607)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2589..2607)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2612..2628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2612..2628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2730..2749)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2741..3550
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2808..2827)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3697..4285
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4186..4205
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggcca caaacggctc caaggtggcc gacggccaga tctccaccga
      721 ggtgtctgag gcccctgtgg ccaatgataa gccaaagaca ctggtggtga aggtgcagaa
      781 gaaggcagca gacctgcctg acagggatac ctggaagggc agattcgatt ttctgatgtc
      841 ttgcgtggga tacgcaatcg gactgggaaa cgtgtggcgg ttcccatacc tgtgcggcaa
      901 gaatggcggc ggcgcctttc tgatccctta cttcctgaca ctgatctttg ccggcgtgcc
      961 actgtttctg ctggagtgca gcctgggcca gtatacctcc atcggaggac tgggcgtgtg
     1021 gaagctggca cccatgttca agggagtggg actggcagca gccgtgctgt ccttttggct
     1081 gaacatctac tatatcgtga tcatctcttg ggccatctac tatctgtaca atagcttcac
     1141 cacaaccctg ccatggaagc agtgcgacaa cccctggaat acagataggt gtttctccaa
     1201 ctattctatg gtgaacacaa ccaatatgac aagcgccgtg gtggagtttt gggagaggaa
     1261 tatgcaccag atgaccgacg gcctggataa gccaggacag atcaggtggc ctctggcaat
     1321 caccctggcc atcgcctgga tcctggtgta cttctgcatc tggaagggcg tgggctggac
     1381 aggcaaggtg gtgtattttt ccgccaccta cccttatatc atgctgatca tcctgttctt
     1441 taggggagtg acactgccag gagcaaagga gggcatcctg ttctacatca cccctaactt
     1501 tagaaagctg tccgactctg aagtgtggct ggatgccgcc acccagatct tcttttccta
     1561 tggactggga ctgggatctc tgatcgccct gggaagctac aactccttcc acaacaacgt
     1621 gtacagggac tctatcatcg tgtgctgtat caattcttgt acaagcatgt tcgccggctt
     1681 tgtgatcttc agcatcgtgg gcttcatggc ccacgtgacc aagcgcagca tcgcagatgt
     1741 ggcagcatcc ggaccaggac tggcctttct ggcataccct gaggcagtga cccagctgcc
     1801 catctcccct ctgtgggcca tcctgttctt ttctatgctg ctgatgctgg gcatcgacag
     1861 ccagttctgc acagtggagg gctttatcac cgccctggtg gatgagtatc ccaggctgct
     1921 gcgcaaccgg agagagctgt tcatcgccgc cgtgtgcatc atctcttacc tgatcggcct
     1981 gagcaatatc acacagggcg gcatctacgt gttcaagctg ttcgactact atagcgcctc
     2041 cggcatgtcc ctgctgttcc tggtgttctt tgagtgcgtg agcatcagct ggttctacgg
     2101 cgtgaacagg ttttatgata atatccagga gatggtgggc tcccgcccat gcatctggtg
     2161 gaagctgtgc tggtctttct ttacccccat catcgtggcc ggcgtgttca tctttagcgc
     2221 cgtgcagatg acaccactga ccatgggcaa ctacgtgttc ccaaagtggg gacagggagt
     2281 gggatggctg atggcactga gctccatggt gctgatcccc ggctacatgg cctatatgtt
     2341 tctgacactg aagggcagcc tgaagcagcg gatccaggtc atggtgcagc cttccgagga
     2401 catcgtgaga ccagagaatg gaccagagca gccacaggca ggatctagca ccagcaagga
     2461 ggcctacatc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2521 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2581 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2641 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2701 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2761 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2821 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2881 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2941 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3001 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3061 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3121 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3181 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3241 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3301 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3361 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3421 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3481 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3541 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3601 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3841 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4261 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4321 cttttgctgg ccttttgctc acatgtt
//
